Protein synthesis and transcriptional inhibitors control N-methyl-N′-nitro-N-nitrosoguanidine-induced levels of APC mRNA in a p53-dependent manner

Aruna S. Jaiswal, Satya Narayan

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

In the present study, we show that treatment of wild-type (p53+/+) mouse embryonic fibroblast (MEF) cells with a DNA-alkylating agent, N-methyl-N′-nitro-N-nitro-soguanidine (MNNG), resulted in increased levels of adenomatous polyposis coli (APC) mRNA compared to p53 gene-knocked out (p53-/-) MEF cells, indicating that p53 is required for APC expression after alkylation damage. By using HCT-116 colon cancer cells (containing wild-type p53 gene) or p53-/- MEF cells transfected with a pCMV-p53 overexpression plasmid [p53-/-(CMV-p53)], we show that p53 is a labile factor for APC gene expression, and that pretreating HCT-116 cells with a protein synthesis inhibitor, cycloheximide (CHX), inhibited MNNG-induced APC mRNA levels by inhibiting p53 protein synthesis. The effect of CHX on p53 protein synthesis was reversible, as the withdrawal of CHX permitted p53 protein synthesis to resume with a concomitant increase in APC mRNA levels after MNNG treatment. To examine whether p53 regulates APC gene expression at the transcriptional level, we treated HCT-116 or p53-/-(CMV-p53) MEF cells with 5,6-dichloro-1-β-D-ribofuranosylbenzamidazole (DRB; a transcriptional inhibitor), before the MNNG treatment. Although treatment of cells with DRB resulted in increased p53 protein levels, that the APC mRNA levels were decreased suggests that p53 may enhance APC gene expression upstream of the transcriptional machinery where DRB interacts. That the withdrawal of DRB, and subsequent MNNG treatment, increased the level of APC mRNA indicated that the binding of DRB to the transcriptional machinery was reversible.

Original languageEnglish (US)
Pages (from-to)733-740
Number of pages8
JournalInternational journal of oncology
Volume13
Issue number4
DOIs
StatePublished - Oct 1998
Externally publishedYes

Keywords

  • 5,6-dichloro-1-β-D-ribofuranosylbenzamidazole
  • Adenomatous polyposis coli
  • Colon cancer
  • Cycloheximide
  • DNA damage
  • mRNA
  • p53

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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